Matrix assisted ink transport

ABSTRACT

Provided is a direct write patterning method utilizing a mixture comprising an ink of choice and an ink carrier matrix. The method involves disposing the mixture on a tip or stamp and transporting the mixture from the tip or stamp on a surface to form a pattern that contains the ink. The method does not require chemical or physical modification of either the tip or stamp or the surface prior to transporting the mixture to the surface. The method can be applied for patterning hard inks such as nanomaterials and crystallized polymers and soft inks such as biomaterials including peptides and proteins. Also provided are related biomaterial and hard ink arrays.

RELATED APPLICATIONS

This application claims priority to U.S. provisional Ser. No. 60/945,164filed Jun. 20, 2007, and also to U.S. provisional Ser. No. 60/929,314filed Jun. 21, 2007, and also to U.S. provisional Ser. No. 61/047,642filed Apr. 24, 2008, all of which are hereby incorporated by referencein their entireties.

STATEMENT ON FEDERAL FUNDING

The present invention was developed with use of federal funding fromNSF-NSEC, Grant No. EEC 0118025; and DARPA-ARD, Grant No. DAAD19-03-1-0065; and NSF Grant No. EEC0647560; and ASAF/AFOSRFA9550-08-1-0124. The federal government reserves rights in theinvention.

BACKGROUND

Nanoscience focuses on elucidating the unique chemical and physicalproperties of nanoscale materials that analogous bulk structures do notpossess (37, 38). Bottom-up and top-down approaches have been used tosynthesize and fabricate such nanoscale materials that are metallic (1,4, 5, 11), magnetic (6, 7), semi-conducting (8, 9), silica-based (18),and carbon-based, such as fullerenes, and carbon nanotubes, (3, 73) withfine control over particle size and shape (74, 36). In the last decade,nanoscale materials have been studied and characterized using a varietyof methods and are becoming better understood.

Nanoscale materials are beginning to be utilized in a growing number ofnovel applications including applications, that rely mainly on theability to arrange nano building blocks (NBBs) into deliberate patternswith controlled feature sizes on surfaces, such as nanocircuitintegration (75), biological micro- and nano-array fabrication (76), andnanoscale sensing (77, 78). Current methods for patterning nano buildingblocks into desired locations usually include the following twosteps: 1) a surface pattern-generation step and 2) a nanoparticleself-assembly step. The first step creates pre-patterns on a surfaceusing photolithography, electron beam lithography (EBL), or focused ionbeam (FIB) lithography (79), while in the second step, nanoparticles areexposed to and further assembled along the pre-patterned areas on thesurface (39). Unfortunately, such surface patterning methods can requireexpensive instrumentation and may be complicated and time-consuming. Forexample, avoiding non-specific binding of nanoparticles to unwantedareas during the second step may be often a very difficult, if notimpossible task. Such problem can be especially prominent at the sub-100nm size regime.

Dip-pen nanolithography (DPN) is a single-step direct writing andreading lithography tool utilized for patterning soft inks, such assmall organic molecules, DNA, and proteins (60), in some cases, at themillimeter and centimeter scale (61, 62). In some cases, it may be moredifficult to directly write hard inks, such as nanoparticles,fullerenes, or crystallized conducting polymers, using DPN due toproblems with obtaining an even coating of such hard inks on an AFM tipand controlling the ink's transport rate. As the result, thenanoparticle patterns may become inconsistent and have uncontrollablefeature sizes. In addition, hard inks may in some cases have a tendencyto dry quickly and agglomerate during the DPN process, which makesextended writing times unachievable (63-68).

Thus, a need exists to develop a single step method for directpatterning of hard inks on a surface that will provide a control overthe patterned feature size and will allow for longer writing times. Inparticular, development of direct patterning methods for protein-basednanostructures is important for researchers working in the areas ofproteomics and theranostics. Such methods would allow generatingmulti-component biological nanostructures of proteins, oligonucleotidesand viruses.

U.S. Pat. No. 7,005,378 describes patterning of metallic precursorsincluding use of polyethylene oxide to facilitate patterning.

The paper “On-Wire Lithography” (Qin et al., Science, vol. 309, Jul. 1,2005, 113-115) describes preparation of gap structures and filling thegap with a mixture of a conductive polymer and polyethylene oxide.

US Patent Publication 2003/0162004 (Mirkin et al., NorthwesternUniversity) describes patterning of sol-gel mixtures comprising blockcopolymers.

US Patent Publication 2004/0142106 (Mirkin et al., NorthwesternUniversity) describes patterning of precursor magnetic materials.

US Patent Publication 2002/0122873 (Mirkin et al., NorthwesternUniversity) describes patterning of magnetic nanoparticles usingmagnetic driving forces.

US Patent Publication 2004/0026007 (Hubert et al., MIT) describesdeposition of nanoparticles.

SUMMARY

The present application describes among other things methods of making,articles, devices, compositions, and methods of using.

One embodiment provides a method comprising: providing a tip, providingan ink disposed at the end of the tip, wherein the ink comprises atleast one matrix and at least one nanomaterial different from thematrix, providing a substrate surface, and transporting the ink from thetip to the substrate surface to form a structure on the surfacecomprising both the matrix and the nanomaterial.

In another example, provided is a method comprising: providing a tip,providing an ink disposed at the end of the tip, wherein the inkcomprises at least one polymer and at least one nanomaterial, providinga substrate surface, and transporting the ink from the tip to thesubstrate surface to form a structure on the surface comprising both thepolymer and the nanomaterial.

One advantage for at least one embodiment is that it allows formingpatterns of inks that may be difficult to pattern. Another advantage forat least one embodiment is that it does not require chemical or physicalmodification of the tip or stamp. In addition, this in many cases doesnot require chemical or physical modification of the substrate surfaceand allows transporting ink molecules to the surface in a fashion thatis independent of the substrate surface's material. In many embodiments,the method allows sub-micron and sub 100-nm patterns of hard inks suchas nanomaterials and biomolecules such as proteins or peptides in adirect write high-throughput manner.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 schematically illustrates patterning of nanomaterials usingmatrix assisted Dip-Pen nanolithography (DPN).

FIGS. 2 (A)-(F) present DPN generated patterns of various polymers on avariety of substrates as well as selected height profiles. (A) is atopographic atomic force microscopy (AFM) image of a pattern ofpolyethylene glycol (PEG) with molecular weight (MW) 8,000 on an Ausubstrate at writing speed of 0.16 μm/s. (B) is an AFM image of apattern of PEG (MW 8,000) on a GaAs substrate at writing speed of 0.022μm/s. (C) is an AFM image of a pattern of polyethylene oxide (PEO) withMW 100,000 on a SiO_(x) substrate at writing speed of 0.05 μm/s. (D) isan AFM image of a pattern of PEO (MW 100,000) on an Au substrate atwriting speed of 0.05 μm/s. (E) is an AFM image of a pattern ofpolyethylene imine (PEI) with MW 10,000 on InAs at 0.6 and 0.3 μm/s. (F)is an AFM image of a pattern of a mixture of PEI (MW 10,000) and 2 nm Aunanoparticles on InAs at 0.6 and 0.3 μm/s.

FIGS. 3 (A) and 3 (B) present height profiles of line patterns of: (A)PEI only; (corresponding topographic AFM image n FIG. 2E) and (B) amixture of 2 nm Au nanoparticles and PEI on InAs substrate(corresponding topographic AFM image in FIG. 2F). FIG. 3(C) is a heightprofile of PEO only line patterns on Au (corresponding AFM topographicimage was shown in FIG. 2D).

FIGS. 4 (A)-(D) present images DPN generated arrays. (A) is atopographic AFM image of PEO arrays at contact time of 64, 32, and 16seconds from top to bottom, respectively. (B) shows a topographic AFMimage of dot arrays deposited using a mixture 2 nm Au nanoparticles andPEO, tip substrate contact time is 64, 32 and 16 seconds from top tobottom respectively. (C) is a topographic AFM image of dot arrays usingdeposited using a mixture of 5 nm Au nanoparticles and PEO,tip-substrate contact times 64, 32, 16, and 8 from top to bottomrespectively, the inset shows a Transmission Electron Microscopy (TEM)image of the dot created by DPN on a TEM grid. (D) shows a topographicAFM image of dot arrays deposited using a mixture 13 nm Au nanoparticlesand PEO, tip substrate contact time is 64, 32 and 16 seconds from top tobottom respectively.

FIGS. 5 (A)-(D) present images of patterns generated by DPN using amixture of 4.7 nm magnetic nanoparticle and PEO. (A) is an AFM image ofdot arrays, tip substrate contact time is 64, 32 and 16 seconds from topto bottom respectively. (B) is an AFM image of diamond shape linearrays, writing speed 0.05 μm/s. (C) is a magnetic force microscopy(MFM) image of larger scale dot arrays. The inset shows a single dotscan (G) is a MFM image of an array of diamond shaped lines created byDPN. The inset shows a single diamond shape line scan.

FIGS. 6 (A)-(D) relate to arrays generated by DPN using a mixture offullerene and PEO. (A) is an AFM image of dot arrays at contact times of16, 8 and 4 seconds from top to bottom respectively. (B) is a heightprofile of the dot arrays of (A). (B) presents line arrays at writingspeed of 0.05, 0.1 and 0.2 μm/s respectively. (C) is a 3-dimensional AFMimage of DPN generated lines crossing through the 500 nm gapnanoelectrode. (F) shows I-V curves of the lines of (E).

FIGS. 7 (A) and (B) are respectively a topographic AFM image (A) and aheight profile (B) of fullerene/PEO dot patterns on Au substrategenerated by DPN. Contact times are 64, 32, and 16 sec from top tobottom of FIG. 7A, respectively. FIG. 7 (C) is a height profile offullerene/PEO line patterns on Au substrate generated by DPN at writingspeeds of 0.05, 0.1, and 0.2 μm/s from left to right, respectively. Thecorresponding AFM topography image was shown in FIG. 6B.

FIG. 8 schematically illustrates generating of protein arrays.

FIGS. 9 (A) and (B) present AFM images of anti-chicken IgG AF 488nanoarrays on Au (A) and silicon (B) surfaces generated by matrixassisted (MA)-DPN.

FIG. 10 shows fluorescence microscopy images of anti-chicken IgG AF 488nanoarrays generated by MA-DPN on silicon substrates.

FIG. 11. (A) Plots showing the relationship of the DPN-generated dotsizes with tip-substrate contact time of selected ink materials, theslopes of the plot reflect the according ink's diffusion constant. (B)Charts showing the change of the ink (anti-ubiquitin) diffusion ratewith the adding of PEO at different ratios. (C) Comparison of thediffusion rate of BSA/PEO and anti-ubiquitin/PEO at ratio of 1:5, thechart shows very close diffusion rate. (D) Charts showing that the inkdiffusion rate of IgG and β-galactosidase can be tuned to be very closeat the ink/PEG ratio of 1:5 and 1:7.5, respectively.

FIG. 12. (A) Fluorescent image of DPN generated dot arrays. The AFM tipswere coated one after another with BSA/PEG (green) andanti-ubiquitin/PEG (red), respectively, both at ratio of 1:5, and bothinks were simultaneously patterned using passive one dimensional A-26AFM tip array. (B) Zoomed-in image of the area within the rectangular in(A), which shows sharp fluorescent signal contrast. (C) and (E), AFMimages of DPN generated nanoarrays containing IgG/PEG (1:5) andβ-galactosidase/PEG (1:7.5), respectively. (D) and (F), fluorescentimages of the nanoarrays in (C) and (E) after incubating with accordingfluorescent labeled antibodies.

FIG. 13. (A) Overview and (B) zoomed-in area of the inkwell that usedfor alternative two ink (BSA/PEG and anti-ubiquitin/PEG) coating. (C)Optical and (D) fluorescent microscopy images of the AFM tip array(A-26) used for multiple-ink patterning by DPN. (E) Overview and (F)zoomed-in area of the inkwell that used for IgG/PEG andβ-galactosidase/PEG coating. Inkwell and tip arrays available fromNanoInk, Inc. (Skokie, Ill.).

DETAILED DESCRIPTION Introduction

Priority U.S. provisional Ser. No. 60/945,164 filed Jun. 20, 2007, andpriority U.S. provisional Ser. No. 60/929,314 filed Jun. 21, 2007, andalso priority U.S. provisional Ser. No. 61/047,642 filed Apr. 24, 2008,are all hereby incorporated by reference in their entireties, includingworking examples, figures, claims, and description of variousembodiments.

Copending application serial No. ______ filed on same day as thisapplication, “Patterning with Compositions Comprising Lipids,” to Mirkinet al., is hereby incorporated by reference in its entirety includingfigures, claims, working examples, and description of other embodiments.

Copending application serial No. ______ filed on same day as thisapplication, “Universal Matrix,” to Mirkin et al., is herebyincorporated by reference in its entirety including figures, claims,working examples, and description of other embodiments.

Nanolithography instruments and accessories, including ink wells and penarrays, for direct-write printing can be obtained from NanoInk, Inc.,Chicago, Ill. DIP PEN NANOLITHOGRAPHY® and DPN® are registered NanoInk,Inc. trademarks.

The following patents and co-pending applications relate to direct-writeprinting with use of for example cantilevers, tips, and patterningcompounds are hereby incorporated by reference in their entirety:

U.S. Pat. No. 6,635,311 issued Oct. 21, 2003 (“Methods UtilizingScanning Probe Microscope Tips and Products Therefor or ProducedThereby”) to Mirkin et al., which describes fundamental aspects of DPNprinting including inks, tips, substrates, and other instrumentationparameters and patterning methods;

U.S. Pat. No. 6,827,979 issued Dec. 7, 2004 (“Methods Utilizing ScanningProbe Microscope Tips and Products Therefor or Produced Thereby”) toMirkin et al., which further describes fundamental aspects of DPNprinting including software control, etching procedures, nanoplotters,and arrays formation.

U.S. patent publication number 2002/0122873 A1 published Sep. 5, 2002(“Nanolithography Methods and Products Produced Therefor and ProducedThereby”), which describes aperture embodiments and driving forceembodiments of DPN printing.

U.S. patent publication 2003/0185967 to Eby et al., published Oct. 2,2003 (“Methods and Apparatus for Aligning Patterns on a Substrate”),which describes alignment methods for DPN printing.

U.S. Pat. No. 7,060,977 to Dupeyrat et al., issued Jun. 13, 2006(“Nanolithographic Calibration Methods”), which describes calibrationmethods for DPN printing.

U.S. Patent Publication 2003/0068446, published Apr. 10, 2003 to Mirkinet al. (“Protein and Peptide Nanoarrays”), which describes nanoarrays ofproteins and peptides;

U.S. Regular patent application Ser. No. 10/307,515 filed Dec. 2, 2002to Mirkin et al. (“Direct-Write Nanolithographic Deposition of NucleicAcids from Nanoscopic Tips”), which describes nucleic acid patterning.

U.S. Patent Publication 2003/0162004 to Mirkin et al. published Aug. 28,2003 (“Patterning of Solid State Features by Direct-WriteNanolithographic Printing”), which describes reactive patterning and solgel inks.

U.S. Pat. No. 6,642,129, issued Nov. 4, 2003, to Liu et al. (“Parallel,Individually Addressible Probes for Nanolithography”).

U.S. Pat. No. 6,737,646, issued May 18, 2004, to Schwartz (“EnhancedScanning Probe Microscope and Nanolithographic Methods Using Same”).U.S. Pat. No. 6,674,074 issued Jan. 6, 2004, to Schwartz (“EnhancedScanning Probe Microscope”).

U.S. Pat. No. 7,098,058 issued Aug. 29, 2006.

U.S. Patent publication 2004/0026681 published Feb. 12, 2004.

U.S. Pat. No. 7,005,378 issued Feb. 28, 2006.

U.S. Patent Publication 2004/0175631 published Sep. 9, 2004.

U.S. Pat. No. 7,034,854 issued Apr. 25, 2006.

U.S. Patent Publication 2005/0009206 published Jan. 13, 2005.

U.S. Patent Publication 2005/0272885 published Dec. 8, 2005.

U.S. Patent Publication 2005/0255237 published Nov. 17, 2005.

U.S. Patent Publication 2005/0235869 published Oct. 27, 2005.

U.S. Patent publication 2006/0040057 to Sheehan et al. (Thermal Controlof Deposition in Dip Pen Nanolithography).

Two dimensional arrays are described in US Patent publication no.2008/0105042 to Mirkin et al., filed Mar. 23, 2007, which is herebyincorporated by reference in its entirety including figures, claims,working examples, and other descriptive embodiments.

In some embodiments, the direct-write nanolithography methods describedherein can be particularly of interest for use in preparing bioarrays,nanoarrays, and microarrays based on peptides, proteins, nucleic acids,DNA, RNA, viruses, and the like. See, for example, U.S. Pat. No.6,787,313 for mass fabrication of chips and libraries; U.S. Pat. No.5,443,791 for automated molecular biology laboratory with pipette tips;U.S. Pat. No. 5,981,733 for apparatus for the automated synthesis ofmolecular arrays in pharmaceutical applications;

Direct write methods, including DPN printing, are described in forexample Direct-Write Technologies, Sensors, Electronics, and IntegratedPower Sources, Pique and Chrisey (Eds), 2002.

Scanning probe microscopy is reviewed in Bottomley, Anal. Chem., 1998,70, 425R-475R. Scanning probe microscopes are known in the art includingprobe exchange mechanisms as described in U.S. Pat. No. 5,705,814(Digital Instruments).

The inventors developed a method of patterning utilizing a mixture thatcomprises a polymer and a nanomaterial. In an embodiment of the method,the mixture is first disposed on a tip or stamp and then transportedfrom the tip or stamp on a substrate surface to form a pattern on thesurface that comprises the ink of choice. The method as applied for DipPen Nanolithography printing is illustrated on FIG. 1.

Ink

Ink can be transported to a surface whether from a tip or a stamp orsome other transport originating surface. The ink can be a compositematerial and can comprise at least two components including at least onepolymer and at least one nanomaterial, the nanomaterial being differentthan the polymer. The ink can be initially formulated with use of asolvent and may further comprise solvent or at least residual solventfor the polymer. In many cases, solvent is removed upon disposing theink at the end of a tip or on a stamp surface. In other cases, the inkyet comprises solvent and is used as a liquid. For example, ink can bedelivered by channels to the end of a tip.

A basic and novel feature can be that the ink consists essentially ofthe polymer and the nanomaterial and is substantially free of componentsthat interfere with transport of polymer and nanomaterial. In somecases, the ink comprises at least at least 70%, or at least 90% byweight polymer and nanomaterial. The ink can comprises less than 30% byweight or less than 10% by weight material which is not polymer ornanomaterial.

Polymer

The ink carrier matrix is usually chosen as any material that can berelatively easily patterned by DPN printing. If a specific feature sizeand particular patterns are desired, the polymer material of the inkcarrier matrix can be any material that can be easily patterned by DPNin a well controlled manner as to provide the desired feature size andpattern when used by itself. Preferably, the polymer ink carrier matrixis selected to be such that it satisfies at least some of the followingcriteria:

1) the polymer ink carrier matrix does not chemically react with eitherthe molecules of the ink or the material of the tip or stamp;

2) a transport rate of the polymer ink carrier matrix is a higher than atransport rate of the ink mixed with the matrix;

3) the polymer ink carrier matrix does not interfere with inherentphysical or biological characteristics of the ink.

The ink carrier matrix can be, for example, a polymer matrix. Thepolymer can be a non-biological polymer. The polymer can be a solublepolymer; it can be a linear polymer having a linear polymer backbone oronly small amount of branching. The polymer can be a copolymer, a blockcopolymer, a random copolymer, a terpolymer, or a branched polymer. Apolymer can be functionalized for crosslinking although in many casesthis is not desired, particularly if the polymer is to removed bysolvent washing.

The polymer can be soluble in both water as well as organic solvent ornon-aqueous solvent.

A polymer forming the polymer matrix can be, for example, polyalkyleneoxide, polyalkylene glycol, or polyalkylene imine. In some embodiments,polyalkylene oxide used as a polymer matrix can be a polyalkylene oxidehaving a molecular weight over 50,000. Yet, in some embodiments, apolyalkylene oxide having a molecular weight of about 50,000 or less canbe used.

In some embodiments, polyethylene oxide (PEO) having a molecular weight(MW) of about 100,000 can be preferred as a material for the polymermatrix. Such a polymer has a low melting temperature and can be easilypatterned by itself using DPN.

In general, PEO does not react with many hard inks or biomaterials andthus does not effect their chemical, biological or physicalcharacteristics. In addition, PEO is soluble in a variety of solventsincluding both hydrophilic and hydrophobic solvents, both aqueous andorganic solvents, both polar and non-polar solvents. The good solubilitymakes PEO compatible with a variety of inks. For example, fullerenes orcarbon nanotubes can be mixed with PEO using toluene as a commonsolvent; magnetic nanoparticles can be mixed with PEO usingdichloromethane as a common solvents; Au nanoparticles or water solubleconducting polymers, such as sulphonated polyaniline (SPAN) or dopedpolypyrrole, can be mixed with PEO using water as a common solvent;quantum dots can be mixed with PEO using hexane as a common solvent;biomolecules such as nucleic acids or proteins can be mixed with PEOutilizing an appropriate biological buffer as a common solvent.Moreover, PEO can be patterned on a variety of substrate surfacesincluding metal surfaces such as Au surface, semiconductor surfaces suchas GaAs or InAs surface or oxide surface such as SiO_(x) surface. Lowermolecular weight PEO, also sometimes called polyethylene glycol, can beused.

The polymer and substrate surface can be adapted so that the polymerdoes not chemisorb to or covalently bond with the surface. Also, thepolymer and the nanomaterial can be adapted so that the polymer is notchemically reactive with the nanomaterial.

Nanomaterial

Nanomaterials can be particulate types of materials having at least onelateral dimension of at least about 100 nm or less, or about 50 nm orless, or about 25 nm or less. The nanomaterial can be for example aspherical material, or a substantially spherical material, or anelongated material. For example, a fullerene for purposes here can beconsidered a substantially spherical material. This lateral dimensioncan be a statistical average for many distinct units or particles. Itcan be for example an average particle diameter for substantiallyspherical particles or an average particle length or width for elongatedparticles. The nanomaterial can be organic or inorganic, hard or soft,flexible or rigid. The nanomaterial can be a non-molecular material. Inpreferred embodiments, the nanomaterial can be for example a metalnanoparticle, a magnetic nanoparticle, or a fullerene nanoparticle.

While the methods described herein can be applied to delivery of a widevariety of ink nanomaterials, in many cases, the ink nanomaterial can bea material that is difficult to pattern by itself, without a polymer asink carrier matrix, using DPN printing for example. For example, thetransport rate may be too slow or the transporting too unreliable.

For instance, the ink of choice can be a hard ink including metalnanoparticles such as Au or Ag silver nanoparticles, semiconductornanoparticles as quantum dots, oxide nanoparticles such as silica oralumina particles, magnetic particles, carbon-based particles such asfullerenes and carbon nanotubes, crystalline polymers includingcrystalline conducting polymers.

The method is not limited to patterning hard inks and can be used alsofor patterning soft inks including biomaterials, biomolecules, orbiological macromolecules such as nucleic acids, DNA, RNA, proteins,peptides, polypeptides, antibodies, and oligo- and polysaccharides.Crystallized conducting polymer can be used.

In an embodiment, the nanomaterial comprises a nanoparticlenanomaterial. The nanomaterial can comprise a nanoparticle comprising anaverage particle size of about 2 nm to about 100 nm, or about 2 nm toabout 25 nm.

In other embodiments, the nanomaterial can be a carbon nanotube, whethersingle, double; or multi-walled. The nanomaterial can comprise ananowire or a nanorod. The nanomaterial can comprise asemiconductor-related material and be for example a quantum dot.

The nanomaterial and substrate surface can be adapted so that thenanomaterial does not chemisorb to or covalently bond with the surface.

Tips and Stamps

The tip embodiment will be further described. The stamp embodiment willalso be further described. Many of the parameters described herein suchas the selection of the patterning compound, surface, and contactconditions can be used for both tip and stamp embodiments. Tips andstamps are used in other technologies besides DPN printing andmicrocontact printing.

Tips known in art of DPN printing can be used. Sharp tips can be usedwhich are characterized by a sharp, pointed end. The tip can be forexample a nanoscopic tip. The tip can be for example a scanning probemicroscope tip or an atomic force microscope tip.

Tips can be engineered to be useful for scanning probe or AFMmeasurements if suitably adapted with for example cantilever andfeedback mechanism. In particular, the tip can be disposed at the end ofa cantilever. The tip can be a hollow tip or a solid tip or a non-hollowtip. The tip can comprise a channel for delivery of the ink mixture.Tips including solid, non-hollow, and hollow tips are further describedin for example U.S. Pat. Nos. 6,635,311 and 6,827,979, as well as2002/0122873, which are herein incorporated by reference in theirentirety. WO 2005/115630 to Henderson et al, published Dec. 8, 2005,also describes an elongated beam with elongated aperture for depositionon surfaces. See also US Patent Publication 2006/0096078 to Bergaud etal. for deposition based on slit or groove technology; see also,Espinosa et al., Small, 1, No. 6, 632-635, 2005 for nanofountain probewriting; Lewis et al., Appl. Phys. Lett., 1999, 75, 2689-2691; Taha etal., Appl. Phys. Lett., 2003, 83, 1041-1043; Hong et al, Appl. Phys.Lett., 2000, 77, 2604-2606; Meister et al., Microelectron. Eng., 2003,67-68, 644-650; Deladi et al., Appl. Phys. Lett., 85, 5361-5363.

Tips can comprise hard inorganic, ceramic materials, or softer organicmaterials. Semiconductor materials can be used. Insulative andconductive materials can be used. Tips known in the art of AFM imaging,for example, can be used including silicon or silicon nitride. Forexample, polymer or polymer-coated tips can be used. See, for example,US Patent Publication No. 2005/0255237 to Zhang et al, which is hereinincorporated by reference in its entirety. Polymer tips and cantileversare described in, for example, Mirkin and Liu, US Patent Publication No.2004/0228962, related to scanning probe contact printing.

The tip disposed on the cantilever can be part of a larger structurecomprising a plurality of tips disposed on a plurality of cantilevers.These can be called multipen structures or parallel pen structures. Forexample, the multipen structure can have over 20, or over 100, or over1,000, or over 10,000, or over 100,000, or over 1,000,000 individualtips. The cantilevers and tips can be adapted for individual actuation,wherein one tip can be raised or lowered independently of another tip.Individual actuation is described in for example U.S. Pat. Nos.6,867,443 and 6,642,129 to Liu et al, which are hereby incorporated byreference in their entirety. Electrostatic or thermal actuation can beused.

Tips can be thermally heated and activated for temperature control. Inparticular, the tip can be heated to effect transport.

Tips can comprise an inorganic surface and tips can be used where theyare not modified after fabrication with an organic material or coating.

In one embodiment, a plurality of tips can be provided comprising inkdisposed at the end of the tip, and transporting ink from the tips tothe substrate surface forms a plurality of structures on the surfacecomprising both the polymer and the nanomaterial.

In addition, stamps can be used including stamps for microcontactprinting can be used. See for example Xia and Whitesides, “SoftLithography,” Angew. Chem. Int. Ed., 1998, 37, 550-575, and referencescited therein, for description of microcontact printing including stamps(pages 558-563). In general, stamps are fabricated for massive parallelprinting using Z direction motion rather than serial motions with fineXY motion. Stamps can comprise a single material or can be formed bymultilayering methods including surface treatments to improve printing.One surface layer can supported which has different properties than thesupport, e.g., stiffer. The stamp can comprise a polymer including anelastomer or a crosslinked rubber, such as, for example, a hydrophobicpolymer, such as a silicone polymer or siloxane polymer, which isadapted for accepting ink but also depositing ink. The stamp can bepatterned to form lines, including straight and curvilinear lines, orcircles or dots.

The stamp can be fabricated to have very small structures, which can bea tip. In addition, surfaces can be used which provide reliefstructures. Here, some areas of the surface rise above other areas ofthe surface, and the ink primarily coats the raised up areas.

One of the advantages of the present method is that it does not requirechemical or physical modification of the tip or stamp. I.e. in someembodiments, the tip or stamp can be an unmodified tip or stamp, i.e. atip or stamp not exposed to chemical or physical modification prior tohaving a mixture comprising an ink and ink carrying matrix beingdisposed on the tip or stamp.

The chemical or physical modification of the tip or stamp is usuallyused in the prior art methods to promote or enhance ink coating to thetip or stamp, to promote or enhance ink adhesion to the tip or stampand/or to promote or enhance ink transport from the tip or stamp to thesubstrate surface. Examples of chemical or physical modification of thetip or stamp include but not limited to base treatment to impart acharged surface of the silicon nitride tip, silinization with amino- ormercaptosilanizing agents, non-covalent modification with smallmolecules or polymeric agents such as polyethyleneglycol (PEG).

Substrate Surface

The substrate surface can be a surface of any substrate although thesurface can be adapted to function with the ink, the polymer, thenanomaterial, and the application at hand. Smother substrates aregenerally preferred for providing pattern's higher resolution. Forexample, the substrate surface can be a surface of an insulator such as,for example, glass or a conductor such as, for example, metal, includinggold. In addition, the substrate can be a metal, a semiconductor, amagnetic material, a polymer material, a polymer-coated substrate, or asuperconductor material. The substrate can be previously treated withone or more adsorbates. Still further, examples of suitable substratesinclude but are not limited to, metals, ceramics, metal oxides,semiconductor materials, magnetic materials, polymers or polymer coatedsubstrates, superconductor materials, polystyrene, and glass. Metalsinclude, but are not limited to gold, silver, aluminum, copper, platinumand palladium. Other substrates onto which compounds may be patternedinclude, but are not limited to silica, silicon oxide SiO_(x), GaAs, InPand InAs.

One of the advantages of the present method is that it does not requirefor a substrate surface to be chemical or physical modified prior totransporting the mixture comprising the ink and the ink carrier matrixto the substrate surface. Accordingly, in some embodiments, thesubstrate surface can be an unmodified substrate surface, i.e. asubstrate surface, which was not chemically or physically modified priorto being patterned.

The chemical or physical medication of the substrate surface is usuallyused in the prior art methods to promote ink transport from the tip orstamp to the substrate surface, to enhance ink adhesion to the substratesurface or to covalently modify the substrate surface. Examples ofphysical or chemical modification of the substrate surface include butnot limited to base treatment of a charged surface of silicon oxide,silanization with amino or mercaptosilinizing agents or modificationwith polymers carrying chemically reactive groups.

Another advantage of the present method that it does not requireprepatterning of the substrate surface.

The substrate can be monolithic or comprise multiple materials includingmultiple layers. In a preferred embodiment, the substrate surface is asemiconductor or metal substrate surface.

The substrate surface can present conductive portions, insulativeportions, or both. The conductive portions can be electrodes forexample. The ink can be transported onto or in between electrodes,establishing contact with electrodes.

Ink Transport

The mixture can be transported from a tip or stamp to a substratesurface in several different ways and is not in particular limited.Known methods in DPN printing and microcontact printing can be used. Forinstance, in scanning probe and AFM-related technology, different modescan be used to have tips interact with surfaces, which include contactmode, non-contact mode and intermittent contact mode or tapping mode.Cantilevers can be oscillated. Known feedback methods can be used forpositioning and alignment the X, Y and Z directions.

The transporting of the mixture from the tip to the surface can becarried out by moving the tip only in the Z direction up and down withrespect to the XY plane of the substrate surface to engage with anddisengage with the surface. A contact time can be used and if contact iswhat activates ink flow then ink flows during the contact time. Themixture delivery can be performed without translating the tip over thesubstrate surface, without moving in the XY plane, and holding the tipstationary. Alternatively, the tip can be translated over the surface,moving in the XY plane. Either the tip can be moved and the surface heldstationary, or the surface can be moved and the tip held stationary.

The transporting can be carried out under conditions such as humidity,temperature, and gaseous atmosphere which provide for a water meniscusbetween the tip and surface. For example, relative humidity can be atleast about 25%, or at least about 40%, or at least about 50%, or atleast about 70%. Conditions can be controlled with use of environmentalchambers. The gaseous atmosphere can be air, an inert atmosphere, anatmosphere with controlled humidity, or with the presence of othervolatile or gaseous compounds such as vapors of organic compounds orvolatile solvents such as alcohols like methanol or ethanol. Conditionscan be selected to not favor a water meniscus including, for example,anhydrous conditions or conditions wherein all reagents and surfaces areselected to be free of water.

The transporting can be done manually or by instrument with computercontrol. Software can be used which can facilitate pattern design,calibration, leveling, and alignment. Calibration methods are describedin for example U.S. Pat. No. 7,060,977 to Cruchon-Dupeyrat et al., whichis hereby incorporated by reference. Alignments methods are describe infor example 2003/0185967 to Eby et al., which is hereby incorporated byreference.

The transporting can be done more than once, repetitively, in either thesame spot or at different locations.

The ink transport can be characterized by an ink transport ratecharacterized from transport of mixtures of the polymer and thenanomaterial. The polymer transport can be characterized by a polymertransport rate. The nanomaterial transport can be characterized by ananomaterial transport rate. The polymer transport rate can be fasterthan the nanomaterial transport rate. Also, the ink transport rate canbe more similar to the polymer transport rate than the nanomaterialtransport rate.

In the present method, a transport rate of the mixture is dominated by atransport rate of the ink carrier matrix's material, such as PEO.Accordingly, a size such as length, width, and/or height of the formedpattern(s) is determined by the transport rate of the ink carriermatrix's material, which can be controlled either via varying humidityas discussed above or by changing a contact time between the tip and thesubstrate surface. The ability to write patterns comprising the ink at arate that can be finely tuned by controlling the transport rater of theink carrier matrix's material, such as PEO, is one of the advantages ofthe present method.

Other Lithographies Besides DPN and Microcontact Printing

Soft lithographic methods including microcontact printing can be used.See for example Xia and Whitesides, “Soft Lithography,” Angew. Chem.Int. Ed., 1998, 37, 550-575, which is hereby incorporated by referencein its entirety. Methods using a patterned elastomeric material as mask,stamp, or mold. Besides microcontact printing, other methods includereplica molding (REM), microtransfer molding (μTM), micromolding incapillaries (MIMIC), and solvent-assisted micromolding (SANIM).

Structure

The structure formed as a result of the ink transport on the surface canbe used as is or treated by additional methods such as heat, light,drying, vacuum, or chemical reaction. Such additional treatment canchemically modify the structure or dry the structure. For example, thepolymer can be crosslinked or annealed and morphologically altered.

The structure can be washed to remove the polymer, or at leastsubstantially most of the polymer.

The structure can be characterized by a lateral dimension such aslength, width, diameter such as for example 1 micron or less, or 500 nmor less, or 300 nm or less, or 100 nm or less, or 50 nm or less.

The structure can be a dot or line, and line can be straight orcurvilinear. Arbitrary shapes can be formed including rings, squares,and triangles.

The structure can have a height which can be for example at least about5 nm, or at least about 10 nm, or at least about 15 nm, or at leastabout 20 nm, or at least about 25 nm. The range can be for example about5 nm to about 100 nm, or about 10 nm to about 50 nm, or about 10 nm toabout 25 nm.

Height can be used to detect the presence of nanomaterial. For example,the structure can have a height which is at least two times, twice, orat least three times, or at least four times, the height compared to astructure substantially identical prepared except without thenanomaterial.

The structure can comprise polymer and the nanomaterial, as well asresidual solvent or moisture. The polymer and the nanomaterial can besubstantially evenly distributed, or they can phase separate.

The methods can be repeated to provide a plurality of structures on thesurface including for example array formation comprising at least two,at least 50, at least 100, at least 500, at least 1,000, or at least50,000 structures on a single surface.

Arrays

The method can be particularly useful for the preparation of nanoarrays,arrays on the submicrometer scale having nanoscopic features when usedwith DIP PEN™ nanolithographic printing. Preferably, a plurality of dotsor a plurality of lines are formed on a substrate. The plurality of dotscan be a lattice of dots including hexagonal or square lattices as knownin the art. The plurality of lines can form a grid, includingperpendicular and parallel arrangements of the lines.

The lateral dimensions of the individual patterns including dotdiameters and the line widths can be, for example, about 2,000 or less,about 1,000 nm or less, about 500 nm or less, about 300 nm or less, andmore particularly about: 100 nm or less. The range in dimension can be,for example, about 1 nm to about 750 nm, about 10 nm to about 2,000 nm,about 10 nm to about 500 nm, and more particularly about 100 nm to about350 nm.

The number of patterns in the plurality of patterns is not particularlylimited. It can be, for example, at least 10, at least 100, at least1,000, at least 10,000, even at least 100,000. Square arrangements arepossible such as, for example, a 10×10 array. High density arrays can bepreferred.

The distance between the individual patterns on the nanoarray can varyand is not particularly limited. For example, the patterns can beseparated by distances of less than one micron or more than one micron.The distance can be, for example, about 300 to about 1,500 microns, orabout 500 microns to about 1,000 microns. Distance between separatedpatterns can be measured from the center of the pattern such as thecenter of a dot or the middle of a line.

The methods described herein can be repeated to provide a plurality ofstructures on the surface which are separated from each other by lessthan a micron.

The method can be also applied for forming patterns of larger scalessuch as micron scale, millimeter scale or centimeter scale. Such largerpatterns can be prepared, for example, utilizing microcontact printingfor transporting the mixture comprising the ink of choice and the inkcarrier matrix from a microcontact printing stamp to the substratesurface.

Arrays of Nano-Building Blocks

The method can be applied for patterning hard inks including but notlimited to metal nanoparticles, such as Au or Ag silver nanoparticles;semiconductor nanoparticles, such as quantum dots; oxide nanoparticles,such as silica or alumina particles; magnetic particles; carbon-basedparticles, such as fullerenes and carbon nanotubes, crystalline polymersincluding crystalline conducting polymers. The method can beparticularly useful for forming hard ink arrays. Such hard ink arrayscomprise a substrate and a plurality of patterns that comprise a hardink of choice and a ink carrier matrix. When the hard ink of choicecomprises carbon based material such as fullerene, the hard ink arraycan serve as an electronic device such as a transistor.

Bioarrays

The method can applied for patterning biomaterials such as nucleicacids, proteins or oligo or polysaccharides. In this case, the mixturecomprises an ink that is a biomaterial of choice and an ink carriermatrix which can be a polymer such as polyalkylene oxide or polyalkyleneimine.

In some embodiments, the biomolecule can comprise various kinds ofchemical structures comprising peptide bonds. These include peptides,proteins, oligopeptides, and polypeptides, be they simple or complex.The peptide unit can be in combination with non-peptide units. Theprotein or peptide can contain a single polypeptide chain or multiplepolypeptide chains. Higher molecular weight peptides are preferred ingeneral although lower molecular weight peptides including oligopeptidescan be used. The number of peptide bonds in the peptide can be, forexample, at least three, ten or less, at least 100, about 100 to about300, or at least 500.

Proteins are particularly preferred. The protein can be simple orconjugated. Examples of conjugated proteins include, but are not limitedto, nucleoproteins, lipoproteins, phosphoproteins, metalloproteins andglycoproteins. Proteins can be functional when they coexist in a complexwith other proteins, polypeptides or peptides. The protein can be avirus, which can be complexes of proteins and nucleic acids, be they ofthe DNA or RNA types. The protein can be a shell to larger structuressuch as spheres and rod structures.

Proteins can be globular or fibrous in conformation. The latter aregenerally tough materials that are typically insoluble in water. Theycan comprise a polypeptide chain or chains arranged in parallel as in,for example, a fiber. Examples include collagen and elastin. Globularproteins are polypeptides that are tightly folded into spherical orglobular shapes and are mostly soluble in aqueous systems. Many enzymes,for instance, are globular proteins, as are antibodies, some hormonesand transport proteins, like serum albumin and hemoglobin.

Proteins can be used which have both fibrous and globular properties,like myosin and fibrinogen, which are tough, rod-like structures but aresoluble. The proteins can possess more than one polypeptide chain, andcan be oligomeric proteins, their individual components being calledprotomers. The oligomeric proteins usually contain an even number ofpolypeptide chains, not normally covalently linked to one another.Hemoglobin is an example of an oligomeric protein.

Types of proteins that can be incorporated into a nanoarray of thepresent invention include, but are not limited to, enzymes, storageproteins, transport proteins, contractile proteins, protective proteins,toxins, hormones and structural proteins. Examples of enzymes include,but are not limited to ribonucleases, cytochrome c, lysozymes,proteases, kinases, polymerases, exonucleases and endonucleases. Enzymesand their binding mechanisms are disclosed, for example, in EnzymeStructure and Mechanism, 2^(nd) Ed., by Alan Fersht, 1977 including inChapter 15 the following enzyme types: dehydrogenases, proteases,ribonucleases, staphyloccal nucleases, lysozymes, carbonic anhydrases,and triosephosphate isomerase. Examples of storage proteins include, butare not limited to ovalbumin, casein, ferritin, gliadin, and zein.

Examples of transport proteins include, but are not limited tohemoglobin, hemocyanin, myoglobin, serum albumin, β1-lipoprotein,iron-binding globulin, ceruloplasmin.

Examples of contractile proteins include, but are not limited to myosin,actin, dynein.

Examples of protective proteins include, but are not limited toantibodies, complement proteins, fibrinogen and thrombin.

Examples of toxins include, but are not limited to, Clostridiumbotulinum toxin, diptheria toxin, snake venoms and ricin.

Examples of hormones include, but are not limited to, insulin,adrenocorticotrophic hormone and insulin-like growth hormone, and growthhormone. Examples of structural proteins include, but are not limitedto, viral-coat proteins, glycoproteins, membrane-structure proteins,α-keratin, sclerotin, fibroin, collagen, elastin and mucoproteins.

Natural or synthetic peptides and proteins can be used. Proteins can beused, for example, which are prepared by recombinant methods.

Examples of preferred proteins include immunoglobulins, IgG (rabbit,human, mouse, and the like), Protein A/G, fibrinogen, fibronectin,lysozymes, streptavidin, avdin, ferritin, lectin (Con. A), and BSA.Rabbit IgG and rabbit anti-IgG, bound in sandwich configuration to IgGare useful examples.Spliceosomes and ribozomes and the like can be used.

A wide variety of proteins are known to those of skill in the art andcan be used. See, for instance, Chapter 3, “Proteins and theirBiological Functions: A Survey,” at pages 55-66 of BIOCHEMISTRY by A. L.Lehninger, 1970, which is incorporated herein by reference.

One of the advantages of the method is that it does not requireprepatterning of the substrate surface with a patterning compound priorto transporting a mixture comprising the protein from the tip to thesurface when forming submicron size patterns, i.e. patterns withfeatures having a lateral dimension of less than about 1 micron, or sub100 nm patterns, i.e. patterns having a lateral dimension of less thanabout 100 nm.

Patterning compounds were used by the prior art methods to improvestability of protein containing submicron or sub 100 nm features.Examples of patterning compounds include a sulfur-containing compoundsuch as, for example, a thiol, polythiol, sulfide, cyclic disulfide, asulfur-containing compound having a sulfur group at one end and aterminal reactive group at the other end, such as an alkane thiol with acarboxylic acid end group. Additional patterning compounds are disclosedin US patent publication 2003/0068446 published Apr. 10, 2003, to Mirkinet. al.

Non-specific binding of proteins to the regions of the substratesurface, can be prevented by covering, or “passivating,” those regionsof the substrate surface that were not exposed to the mixture comprisingthe biomolecule and the ink carrier matrix with one or more passivatingcompounds. Known passivating compounds can be used and the invention isnot particularly limited by this feature to the extent that non-specificadsorption does not occur. A variety of passivating compounds can beused including, for example, surfactants such as alkylene glycols whichare functionalized to adsorb to the substrate. An example of a compounduseful for passivating is 11-mercaptoundecyl-tri(ethylene glycol).Proteins can have a relatively weak affinity for surfaces coated with11-mercaptoundecyl-tri(ethylene glycol) and therefore do not bind tosuch surfaces. See, for instance, Browning-Kelley et al., Langmuir 13,343, 1997; Waud-Mesthrige et al., Langmuir 15, 8580, 1999;Waud-Mesthrige et al., Biophys. 1 80 1891, 2001; Kenseth et al.,Langmuir 17, 4105, 2001; Prime & Whitesides, Science 252, 1164, 1991;and Lopez et al., J. Am. Chem. Soc. 115, 10774, 1993, which are herebyincorporated by reference. However, other chemicals and compounds, suchas bovine serum albumin (BSA) and powdered milk, that can be used tocover a surface in similar fashion to prevent non-specific binding ofproteins to the substrate surface. BSA, however, can provide lessperformance than 11-mercaptoundecyl-tri(ethylene glycol). Afterpassivation, the resultant array can be called a passivated array ofproteins or peptides.

After passivation, the matrix can be washed away from the patternedregions on the surface. The use of polyalkylene oxide as the matrixallows retaining the biological activity of the biomaterial in thepatterned regions upon washing away the matrix.

One embodiment of making protein array according to the method isillustrated in FIG. 8.

Applications

Biological, diagnostic, assays, sensors, semiconductor, electronic,photomask repair, transistor fabrication and repair, including fieldeffect transistors, flat panel display fabrication and repair, andmagnetic applications can be benefited with use of the variousembodiments described herein.

Many applications of DPN printing are described in Ginger, Zhang, andMirkin, “The Evolution of Dip Pen Nanolithography,” Angew. Chem. Int.Ed., 2004, 43, 30-45, which is hereby incorporated by reference in itsentirety.

Applications for microcontact printing are described in for example Xiaand Whitesides, “Soft Lithography,” Angew. Chem. Int. Ed., 1998, 37,550-575, and references cited therein, which is hereby incorporated byreference in its entirety. Biological applications include assays,diagnostics, sensor, protein microarrays, nucleic acid and DNAmicroarrays, nanoarrays, cell adhesion and growth, and the like.Biodiagnostic applications are described in for example Rose & Mirkin,“Nanostructures in Biodiagnostics,” Chem. Rev., 2005, 105, 1547-1562,which is hereby incorporated by reference in its entirety. DNAmicroarrays are described in DNA Microarrays, A Practical Approach, Ed.Schena, 1999, Oxford University Press. Applications for protein andpeptide nanoarrays are described in for example US Patent PublicationNo. 2003/0068446 to Mirkin et al., which is hereby incorporated byreference in its entirety. For example, surfaces can be patterned withcompounds adapted for capturing a variety of proteins and peptidestructures.

Further assays can be developed including for example testing fordiseases such as HIV. See for example Lee et al, “Nano-Immunoassays forUltrahigh Sensitive/Selective Detection of HIV,” NanoLett. 2004, 4,1869-1872, which is hereby incorporated by reference in its entirety.This describes patterning of MHA, which is then deprotonated so featuresare negatively charged. Monoclonal antibodies to the HIV-1 p24 antigenare then immobilized on the MHA and then exposed to plasma samples takenfrom infected patients. Nanoparticle probes can be used to detect andamplify the signal.

In these and other biological applications, surfaces can be passivatedto prevent non-specific binding including non-specific protein binding.See also US Patent Publication No. 2005/0009206 to Mirkin et al, whichis hereby incorporated by reference in its entirety.

In field effect transistor applications, sources, drains, gates,electrodes, and channels can be fabricated by methods known in the arts.

The invention is further illustrated by, though in no way limited to,the following working examples.

WORKING EXAMPLES 1. Materials and Instrumentation

Polyethylene oxide (PEO, MW=100,000), polyethylene glycol (PEG,MW=8,000), and polyethyleneimine (PEI, MW=2,000) were purchased fromSigma-Aldrich (Milwaukee, Wis.). Au nanoparticles (AuNP) solutions wereobtained from Ted Pella (Redding, Calif.). Magnetic nanoparticles (MNP)were synthesized.

Fullerene was purchased from Mer Corporation (Tucson, Ariz.).Acetonitrile, dichloromethane, toluene were purchased from FisherScientific (Fairlawn, N.J.). All chemicals were used as received.

Si/SiO_(x) wafer with 500 nm oxide coating layer were purchased fromWaferNet, Inc. (San Jose, Calif.). Gold substrates were obtained bythermal evaporation of a gold thin film (30 nm) on a Si/SiO_(x)substrate pre-coated with a Ti adhesion layer (7 nm). GaAs and InAswafers were purchased from Wafer World Inc. (West Palm Beach, Fla.).

All DPN experiments were performed on a ThermoMicroscopes CP AFM (VeecoInstruments Inc., CA), which was enclosed in a humidity-controlledchamber and driven by commercially available DPN software (NanoInk Inc.,Chicago, Ill.). The humidity was controlled at 70% for all PEO relatedexperiments, and 50% for PEI experiments. AFM probes (S-1 or S-2) werepurchased from NanoInk Inc., with spring constants of 0.041 N/m and 0.1N/m, respectively. MFM data were obtained with a DI multimode SPM (VeecoInstruments Inc., CA), using a pre-magnetized AFM probe.

Preparation of Inks

For all DPN experiments, PEO and PEG solutions (16 mg/mL) were made bydissolving PEO in acetonitrile, dichloromethane, water, or toluene. Toprepare the AuNP/PEO ink, PEO (16 mg/mL) in acetonitrile was mixed witha AuNP solution at a volume ratio of 1:1 (2 nm AuNP), 2:1 (5 nm AuNP),and 4:1 (13 nm AuNP). To prepare the 4.7 nm MNP/PEO solution, PEO (16mg/mL) in dichloromethane was mixed with a MNP solution at a volumeratio of 2:1. To prepare the fullerene/PEO ink, PEO (16 mg/ml) intoluene was mixed with a saturated fullerene solution in toluene at avolume ratio of 1:2. To prepare the 2 nm AuNP/PEI ink, a 5% diluted PEIwater solution was mixed with a 2 nm AuNP solution at the volume ratioof 1:1.

2. Matrix-Assisted DPN of Nanobuilding Blocks A. Polymer Only Controls

FIG. 2 shows control patterns of polyethylene glycol (PEG, MW 8,000),polyethylene oxide (PEO, MW 100,000), and polyethylene imine (PEI, MW2000) created using DPN on several types of substrates. In particular,FIG. 2A and FIG. 2B present topographic AFM images of DPN-generated PEGpatterns on Au (writing speed of 0.16 μm/s) and GaAs (writing speed of0.022 μm/s), respectively. FIG. 2C and FIG. 2D show DPN-generated PEOpatterns on SiO_(x) and Au respectively with writing speed of 0.05 μm/sfor both. FIG. 2E demonstrates direct patterning of PEI with writingspeeds of 0.6 and 0.3 μm/s on an InAs substrate. The correspondingheight profile in FIG. 3A shows that different writing speeds result indifferent pattern heights. The faster writing speed (0.6 μm/s) producessmaller height (1.75 nm), while the slower writing speed (0.3 μm/s)produces bigger height (2.75 nm).

FIG. 2F demonstrates the ability of PEI to act as a carrier matrix bypresenting DPN patterns of a mixture of PEI and 2 nm Au nanoparticles onan InAs substrate produced with writing speeds of 0.1 and 0.05 μm/s. Thecorresponding height profiles in FIG. 3B demonstrate that 0.1 μm/swriting speed produces pattern having height of 12 nm, while 0.05 μm/swriting speed produces pattern having height of 14 nm. Comparison of theheight profiles demonstrates that the pattern of the mixture containing2 nm Au nanoparticles is distinctly greater than that of PEI only. Thisindicates the presence of Au nanoparticles in the patterns prepared fromthe mixture containing Au nanoparticles.

B. Au Nanoparticles

The capability of these polymers to act as a carrier matrices wasdemonstrated for common nanomaterials. Specifically, FIG. 4 shows arraysof Au nanoparticles (AuNP) generated using direct single-step patterningprocess. As a control, FIG. 4A shows a topographic AFM image of dotarrays produced using PEO only, with tip-substrate contact times of 64,32, and 16 seconds from top to bottom respectively. The feature heightsof the obtained dot arrays are 8.5, 3.3, and 1.7 nm for contact times of64, 32, and 16 seconds respectively, see TABLE 1. FIG. 4A, FIG. 4B andand FIG. 4C are topographic AFM images of DPN generated dot arrays of 2,5, and 13 nm Au nanoparticles mixed with PEO respectively. TABLE 1 liststhe heights of these structures. Clearly, all of the nanoscale featurescontaining Au nanoparticles are much greater in height than those ofonly PEO. The height increase is larger for patterns containingnanoparticles of bigger diameters. In a similar manner, a mixture of 5nm Au nanoparticles and PEO was patterned on a Transmission ElectronMicroscope (TEM) grid. The inset of FIG. 3E, which is a TEM image of aDPN generated dot on the TEM grid, demonstrates clusters of Aunanoparticles, which proves the presence of Au nanoparticles in thesepatterns.

TABLE 1 Heights of DPN generated dot features, nm AuNP/ AuNP/ AuNP/Contact PEO MNP/ PEO PEO PEO C₆₀/ time, s only PEO 2 nm 5 nm 13 nm PEO64 8.5 27.4 20.8 25.8 32.3 21.8 32 3.3 23.1 13.8 16.1 23.5 14.6 16 1.718.3 8.6 10.6 18.5 9.8

C. Magnetic Nanoparticles

Patterns of magnetic nanoparticles (MNP) were also created using amatrix-assisted DPN. FIG. 5 features the patterns containing 4.7 nmmagnetic nanoparticles (MNP) prepared using PEO as a carrier matrix.FIG. 5A and FIG. 5B are topography AFM images of DPN-generated dotarrays with tip substrate contact of 64, 32, and 16 sec from top tobottom, and diamond-shape line patterns at writing speed of 0.05 μm/s,respectively. Again, an obvious height difference was observed whencomparing the heights of these patterns with those of pure PEO, seeTABLE 1. The increased height for patterns prepared from mixturescontaining MNPs indicates the MNPs are embedded in these patterns.

To further prove the presence of the MNPs inside patterns prepared froma mixture containing MNPs, the patterns were further characterized usingMagnetic Force Microscopy (MFM), a technique which shows clear contrastbased on the magnetism of the sample. In the MFM images in FIG. 5C andFIG. 5D, the patterned features containing MNPs can be undoubtedlydistinguished from the non-magnetic bare SiO_(x) substrate. This strongcontrast even is observed for a single feature, see insets in FIG. 5Cand FIG. 5D indicating that magnetic particles were evenly distributedthroughout the entire patterned feature. The MFM image of a single linepattern, see inset in FIG. 5D shows magnetic clusters inside thepattern. These kinds of clusters are not observed in patterns of purePEO. This observation indicates that these clusters are pockets of MNPs.The large area patterns presented in FIG. 4 (C)-(D) also demonstratethat the matrix assisted DPN can provide extended writing times as wellas smooth and well-controlled ink transfer rate.

D. Fullerenes

In addition to Au nanoparticles and magnetic nanoparticles, DPN patternsof carbon-based nanomaterials (fullerenes) were also generated using PEOas a carrier. The ability to pattern fullerenes is particularlyimportant due to their potential application in nanoelectronics (71).

FIG. 6 shows DPN-generated nanoarrays of a mixture of fullerene and PEO.FIG. 6A shows a dot array with tip-substrate contact times of 16, 8, and4 s (top to bottom). 80 nanometer feature sizes were easily created atthe 4 second contact time (FIG. 6A), proving that sub-100 nm featurescan be obtained easily using this technique. With contact times of 64,32, and 16 s, features of 21.8, 14.6 and 9.8 nm in height were produced,see TABLE 1 and a topography AFM image and corresponding height profilein FIG. 7A and FIG. 7B. Again, these heights are greatly increasedcompared to those of the corresponding pure PEO patterns, indicative ofthe presence of fullerenes in the DPN dot arrays generated from themixture containing fullerenes. These same trends regarding heightincreases, see FIG. 7C, were observed for continuous lines producedusing the mixture of fullerene and PEO (writing speeds=0.05, 0.1, and0.2 μm/s), see FIG. 6B.

As a proof-of-concept, as well as to further confirm that fullerenemolecules indeed are patterned in these DPN-generated features, thefirst fullerene-based transistor was built via DPN. Lines of thefullerene/PEO ink were generated across an EBL-generated nanoelectrodewith a gap size of 500 nm. The 3D topographic AFM image in FIG. 6Cclearly shows two crossed, continuous lines wired across these gaps.Current-voltage (I-V) measurements monitoring the output current of thisdevice at voltages ranging from −0.7 V to 0.85 V are shown in FIG. 6D.The black line is a plot of the I-V response of the transistor measuredin a dark environment, while the red (gray) line shows the currentobtained under illumination with a Xe lamp (150 W). The observedincrease in current (˜6 times more, ˜0.015 pA at 0.85 V vs. ˜0.10 pA at0.85 V) is a characteristic response of fullerene molecules to lightillumination (70, 72). Such a response indicates that the photoactivefullerene molecules are present in an active state inside theDPN-generated patterns. In addition, the precise delivery offullerene/PEO lines within the 500 nm gapped nanoelectrode alsodemonstrates a high spatial resolution of DPN.

3. Protein Nanoarrays

Nanoarrays of goat anti-chicken IgG Alexafluor 488 were prepared by amatrix assisted DPN as illustrated in the general scheme presented inFIG. 8. A low molecular weight polymer (poly-ethylene glycol, MW=8000)was used as a matrix to transport anti-chicken IgG AF 488 from the AFMtip to the substrate surface. PEG is an excellent material to resistnon-specific protein adsorption on surfaces. The use of PEG as a matrixallows one to wash away PEG after generating protein nanoarray to retainthe biological activity of the protein. DPN was performed at a relativehumidity of 75% and at 25° C. Unmodified NanoInk type A tips were dipcoated with a mixture containing the antibody and PEG and dried withnitrogen. FIG. 9A and FIG. 9B demonstrate AFM images of generatednanoarrays of anti-chicken IgG Alexafluor 488 by MA-DPN method on goldand silicon substrates, respectively. The anti-chicken IgG Alexafluor488 nanoarrays were further characterized by fluorescence microscopy asshown FIG. 10.

The AFM and fluorescence images clearly indicate that one can generateuniform nanoarrays of proteins using MA-DPN. The matrix encapsulatedproteins are shown to be biologically active as indicated by our resultswith microarrays generated by microcontact printing.

ADDITIONAL EXAMPLES

A significant application of this universal ink is the capability ofsimultaneous patterning of multiple biomolecules, and the retaining oftheir bioactivities. As stated previously, each ink has its owndiffusion rate, which makes it extremely difficult (if possible) forsimultaneous patterning of multiple inks, and further for feature sizecontrol via the tip-substrate contact time. FIG. 11A shows the inkdiffusion rate of PEG as well as four biomolecules in PBS buffer. Onecan easily see that the ink diffusion rate varies dramatically accordingto different ink materials selected, which will sequentially become amajor issue if we anticipate very similar or identical feature sizeduring simultaneous multiple ink patterning. For example, the slope ofpure IgG can be as high as 30.81, while that of anti-ubiquitin is only11.30, which means at the same tip-substrate contact time (4 sec), thegenerated dot size will be 439.0 nm for β-galactosidase and 144.7 nm forBSA, which indeed varies a lot. What is more, the different slopes alsomeans that the increase trend of the dot size is also different.

However, using the universal ink where PEG works as an ink carrier, theink diffusion rate can be easily tuned within a certain range. In orderto prove this point, we have monitored the ink diffusion rate change ofthe mixture of anti-ubiquitin/PEG at different ratios (FIG. 11B). Atanti-ubiquitin:PEG ratio of 1:2, the diffusion rate of the mixed inkjumps to 28.72 from 11.30, and it further increases to 29.41 at theratio of 1:5. Plots in FIGS. 11C and 11D not only give more examples ofsuch capability PEG has, but also show that the diffusion rate of eachindividual ink can be tuned within certain range, and what is more, wecan make two different inks have very similar diffusion rate. This is animportant parameter that facilitates the precise control of each ink'sfinal feature size and the sequential size increase trend aftermultiple-ink DPN patterning since the tip-substrate contact time willalways be the same (as the AFM probe array we used is a passive mode).Except the ink carrier capability, another important role PEG plays inthe universal ink kit is its ability to tune the ink's diffusion rate.

One then used one dimensional AFM tip array (Model No.: A-26, NanoInkInc., Skokie, Ill.) for simultaneous multiple ink patterning via DPN.Two composite inks containing fluorescent labeled BSA (green color) andanti-ubiquitin (red color), were coated in every other AFM probes,respectively, using the inkwell (NanoInk Inc., Skokie, Ill.) thatspecially designed for such purposes. Both the optical microscopy imagesof the inkwell we used and the AFM tip arrays before and afterink-coating are shown in FIG. 13. The diffusion rates of the two inkswere intentionally tuned very similar following the ratio of 1:5 forboth BSA:PEG and anti-ubiquitin:PEG shown in FIG. 11C. DPN was doneunder the same experimental conditions as described in FIG. 11C. Thefluorescent images in FIG. 12A clearly proved that two different kindsof biomolecules (BSA in green and anti-ubiquitin in red) weresimultaneously patterned into designed array. The zoomed-in image inFIG. 12B shows more details and clear contrast of the fluorescentsignal. In order to compare the variation of generated pattern sizes,one took AFM images after DPN experiment to characterize the generateddot sizes. As a representative, at tip-substrate contact time of 32 sec,the average dot diameter is 328.3 nm for BSA and 306.1 nm foranti-ubiquitin, which has only less than 7% variation (AFM images notshown). On the other side, the generated dot sizes would be 284.3 nm and223.1 nm if not mixed with PEG based on the plots shown in FIG. 11A.

To further prove the bioactivities of the patterned biomolecules, wefirst generated IgG and β-galactosidase patterns individually. FIGS. 12Cand 12E are AFM images of generated IgG and β-galactosidase dot arraysat tip-substrate contact time of 32 sec. The average dot diameter is347.2 nm for IgG and 380.3 nm for β-galactosidase, which has around 8%variation. Similarly, the generated biomolecular dot sizes would be251.0 nm and 439.1 nm, respectively, if without PEG according to FIG.11A.

One then incubated the biomolecular arrays into according antibodybuffer solution. The according fluorescent images in FIGS. 12D and 12Findicate that both anti-IgG (green) and anti-β-galactosidase (red) cansuccessfully bind on the pre-generated dot arrays of antigen molecules,which means the patterned IgG and β-galactosidase still remain theirbioactivities.

All of the publications, patent applications and patents cited in thisspecification are incorporated herein by reference in their entirety.

LIST OF REFERENCES

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1. A method comprising: providing a tip, providing an ink disposed atthe end of the tip, wherein the ink comprises at least one polymer andat least one nanomaterial, providing a substrate surface, andtransporting the ink from the tip to the substrate surface to form astructure on the surface comprising both the polymer and thenanomaterial.
 2. The method of claim 1, wherein the tip is a nanoscopictip.
 3. The method of claim 1, wherein the tip is a scanning probemicroscopic tip.
 4. The method of claim 1, wherein tip is an atomicforce microscopic tip.
 5. The method of claim 1, wherein the tip is anon-hollow tip.
 6. The method of claim 1, wherein the tip is a hollowtip.
 7. The method of claim 1, wherein the tip comprises an inorganicsurface.
 8. The method of claim 1, wherein the tip is not surfacemodified with an organic material.
 9. The method of claim 1, wherein aplurality of tips are provided comprising ink disposed at the end of thetip, and transporting the ink from the tips to the substrate surfaceforms a plurality of structures on the surface comprising both thepolymer and the nanomaterial.
 10. The method of claim 1, wherein the tipis heated to effect transport.
 11. The method of claim 1, wherein thetip is an actuated tip.
 12. The method of claim 1, wherein the tip isdisposed at the end of a cantilever.
 13. The method of claim 1, whereinthe nanomaterial comprises a nanoparticle nanomaterial.
 14. The methodof claim 1, wherein the nanomaterial comprises a nanoparticle comprisingan average particle size of about 2 nm to about 100 nm.
 15. The methodof claim 1, wherein the nanomaterial comprises a nanoparticle comprisingan average particle size of about 2 nm to about 25 nm.
 16. The method ofclaim 1, wherein the nanomaterial comprises a substantially sphericalmaterial or an elongated material.
 17. The method of claim 1, whereinthe nanomaterial comprises a metal nanoparticle, a magneticnanoparticle, or a fullerene nanoparticle.
 18. The method of claim 1,wherein the nanomaterial comprises a carbon nanotube.
 19. The method ofclaim 1, wherein the nanomaterial comprises a nanowire or a nanorod. 20.The method of claim 1, wherein the nanomaterial comprises a quantum dot.21. The method of claim 1, wherein the nanomaterial comprises at leastone biological macromolecule.
 22. The method of claim 1, wherein thenanomaterial comprises at least one biomolecule.
 23. The method of claim1, wherein the nanomaterial comprises at least one protein.
 24. Themethod of claim 1, wherein the nanomaterial comprises at least oneantibody.
 25. The method of claim 1, wherein the nanomaterial comprisesat least one crystallized conducting polymer.
 26. The method of claim 1,wherein the polymer is a non-biological polymer.
 27. The method of claim1, wherein the polymer is a synthetic, linear polymer.
 28. The method ofclaim 1, wherein the polymer is a soluble polymer.
 29. The method ofclaim 1, wherein the polymer is soluble in water and organic solvent.30. The method of claim 1, wherein the polymer is a poly(alkylene oxide)or a poly(alkylene imine).
 31. The method of claim 1, wherein thepolymer is polyethylene oxide having a molecular weight of more than50,000.
 32. The method of claim 1, wherein the ink consists essentiallyof the polymer and the nanomaterial.
 33. The method of claim 1, whereinthe ink further comprises a solvent for the polymer.
 34. The method ofclaim 1, wherein the polymer is not covalently bound or chemisorbed tothe nanomaterial.
 35. The method of claim 1, wherein the polymer doesnot chemisorb to or covalently bond with the surface.
 36. The method ofclaim 1, wherein the nanomaterial does not chemisorb to or covalentlybond to the surface.
 37. The method of claim 1, wherein the polymer isnot chemically reactive with the nanomaterial.
 38. The method of claim1, wherein the substrate surface is a semiconductor or metal substratesurface.
 39. The method of claim 1, wherein the substrate surfacecomprises a nanoelectrodes gap.
 40. The method of claim 1, wherein thetransporting is carried out under humidity and environmental conditionsproviding for a meniscus between the tip and the surface.
 41. The methodof claim 1, wherein the transporting is carried out with at least 40%relative humidity.
 42. The method of claim 1, wherein the transportingis carried out with at least 70% relative humidity.
 43. The method ofclaim 1, wherein the structure has a lateral dimension of about 1 micronor less.
 44. The method of claim 1, wherein the formed pattern ischaracterized by a lateral dimension of about 100 nm or less.
 45. Themethod of claim 1, wherein the structure is a dot or a line.
 46. Themethod of claim 1, wherein the structure has a height of at least 10 nm.47. The method of claim 1, wherein the structure has a height which isat least twice the height compared to a structure substantiallyidentically prepared except without the nanomaterial.
 48. The method ofclaim 1, wherein the structure has a height which is at least threetimes the height compared to a structure substantially identicallyprepared except without the nanomaterial.
 49. The method of claim 1,wherein the structure has a height which is at least four times theheight compared to a structure substantially identically prepared exceptwithout the nanomaterial.
 50. The method of claim 1, wherein thestructure comprises the polymer and nanomaterial substantially evenlydistributed.
 51. The method of claim 1, wherein the polymer ischaracterized by a transport rate, and the nanomaterial is characterizedby a transport rate, and the polymer transport rate is faster than thenanomaterial transport rate.
 52. The method of claim 1, wherein the inkis characterized by an ink transport rate, the polymer is characterizedby a polymer transport rate, and the nanomaterial is characterized by ananomaterial transport rate, and wherein the ink transport rate is moresimilar to the polymer transport rate than the nanomaterial transportrate.
 53. The method of claim 1, wherein method is repeated to provide aplurality of structures on the surface.
 54. The method of claim 1,wherein method is repeated to provide a plurality of structures on thesurface which are separated from each other by less than a micron. 55.The method of claim 1, wherein the transporting is carried out bycontacting the tip with the surface and holding the tip stationary. 56.The method of claim 1, wherein the transporting is carried out bycontacting the tip with the surface and moving the tip with respect tothe surface, or moving the surface with respect to the tip.
 57. Themethod of claim 1, wherein the transporting is carried out in a tappingmode.
 58. The method of claim 1, further comprising the step of removingat least some of the polymer from the structure.
 59. The method of claim1, wherein the tip is a nanoscopic tip, the polymer is a solublepolymer, and the nanomaterial is a nanoparticle.
 60. The method of claim1, wherein the tip is a scanning probe tip, the polymer is a syntheticpolymer, and the nanomaterial is a nanoparticle, a protein, or anantibody.
 61. The method of claim 1, wherein the tip is an AFM tip, thepolymer is a polyethylene oxide, polyethylene glycol, or polyethyleneimine, and the nanomaterial is a nanoparticle or a biological material.62. A method comprising: providing an elastomeric, patterned stamp,providing an ink disposed on the surface of the stamp, wherein the inkcomprises at least one polymer and at least one nanomaterial, providinga substrate surface, and transporting the ink from the stamp to thesubstrate surface to form a structure on the surface comprising both thepolymer and the nanomaterial.
 63. A method comprising: providing a tipor an elastomeric, patterned stamp, providing an ink disposed on thesurface of tip or the stamp, wherein the ink comprises at least onepolymer and at least one nanomaterial, providing a substrate surface,and transporting the ink from the tip or the stamp to the substratesurface to form a structure on the surface comprising both the polymerand the nanomaterial.
 64. A method comprising (A) providing a tip orstamp; (B) providing a mixture comprising an ink and a carrier matrix,wherein the carrier matrix is selected from a) polyalkylene oxideshaving a molecular weight of more than 50,000 and b) polyalkyleneimines; (C) disposing the mixture at the tip or stamp; (D) providing asubstrate surface; and (E) transporting the mixture from the tip orstamp to the substrate surface to form a pattern on the substratesurface such that the pattern comprises the ink.
 65. The method of claim64, wherein the tip or stamp is a chemically or physically unmodifiedtip or stamp.
 66. The method of claim 64, wherein the tip or stamp is atip.
 67. The method of claim 64, wherein the tip is a scanning probemicroscopic tip.
 68. The method of claim 64, wherein the tip is anatomic force microscopic tip.
 69. The method of claim 64, wherein thedisposing comprises immersing the tip in the mixture.
 70. The method ofclaim 64, wherein the disposing comprises immersing the tip in themixture and drying the mixture.
 71. The method of claim 64, wherein thetip or stamp is a microcontact printing stamp.
 72. The method of claim64, wherein the ink is a hard ink.
 73. The method of claim 64, whereinthe ink is a hard ink and the hard ink is selected from the groupconsisting of nanoparticles, carbon based materials and crystallizedpolymers.
 74. The method of claim 64, wherein the ink is a hard ink andthe hard ink is selected from the group of metal nanoparticles, magneticnanoparticles and fullerenes.
 75. The method of claim 64, wherein theink comprises at least one biomolecule.
 76. The method of claim 75,wherein the biomolecule is selected from the group consisting of nucleicacids, peptides and proteins.
 77. The method of claim 64, wherein theink comprises at least one protein.
 78. The method of claim 77, whereinsaid at least one protein is an antibody.
 79. The method of claim 64,wherein the polymer is polyethylene oxide.
 80. A method comprising (A)providing a tip or stamp; (B) providing a mixture comprising a hard inkand a carrier matrix; (C) disposing the mixture on the tip or stamp; (D)providing a substrate surface; and (E) transporting the mixture from thetip or stamp to the substrate surface to form a pattern on the substratesurface such that the pattern comprises the hard ink.
 81. An hard inknanoarray comprising (A) a substrate and (B) a plurality of patterns onthe substrate, the patterns comprising a hard ink material and a matrixmaterial.
 82. A method comprising (A) providing a tip or stamp; (B)providing a mixture comprising an ink and a carrier matrix, wherein theink comprises at least one biomolecule and the carrier matrix comprisesa material selected from the group consisting of polyalkylene oxides andpolyalkylene imines; (C) disposing the mixture at the tip or stamp; (D)providing a substrate surface; (E) transporting the mixture from the tipor stamp to the substrate surface to form at least one pattern on thesubstrate surface such that the at least one pattern comprises the atleast one biomolecule.
 83. A method comprising (A) providing a tip orstamp; (B) providing a mixture comprising an ink and a matrix such thata transport rate of the matrix is greater than a transport rate of theink; (C) disposing the mixture on the tip or stamp; (D) providing asubstrate surface; and (E) transporting the mixture from the tip orstamp to the substrate surface to form at least one pattern on thesubstrate surface such that the at least one pattern comprises the ink.84. A method comprising: providing a tip, providing an ink disposed atthe end of the tip, wherein the ink comprises at least one matrix and atleast one nanomaterial different from the matrix, providing a substratesurface, and transporting the ink from the tip to the substrate surfaceto form a structure on the surface comprising both the matrix and thenanomaterial.
 85. The method of claim 84, wherein the matrix is apolymer.
 86. The method of claim 1, wherein the tip is part of a largerstructure comprising a plurality of tips with inks disposed at the endsof the tips, wherein the inks comprise multiple biomolecules which aresimultaneously transported from the tips to the substrate.
 87. Themethod of claim 1, wherein the tip is part of a larger structurecomprising a plurality of tips with inks disposed at the ends of thetips, wherein the inks comprise multiple proteins.
 88. The method ofclaim 1, wherein the tip is part of a larger structure comprising aplurality of tips with inks disposed at the ends of the tips, whereinthe inks are characterized by a diffusion rate which is tuned so that atleast two different inks have similar diffusion rate during transport.89. The method of claim 1, wherein the tip is part of a larger structurecomprising a plurality of tips with inks disposed at the ends of thetips, wherein the inks comprise multiple biomolecules which aresimultaneously transported from the tips to the substrate, and the ratiobetween polymer and biomolecule is different on different tips.
 90. Themethod of claim 1, wherein the tip is part of a larger structurecomprising a plurality of tips with inks disposed at the ends of thetips, wherein the inks comprise multiple biomolecules which aresimultaneously transported from the tips to the substrate to form dotswhich have an average dot diameter characterized by less than 7%variation.
 91. The method of claim 1, wherein the tip is part of alarger structure comprising a plurality of tips with inks disposed atthe ends of the tips, wherein the inks comprise multiple proteins whichare simultaneously transported from the tips to the substrate, and theratio between polymer and polymer is different on different tips. 92.The method of claim 1, wherein the tip is part of a larger structurecomprising a plurality of tips with inks disposed at the ends of thetips, wherein the inks comprise multiple proteins which aresimultaneously transported from the tips to the substrate to form dotswhich have an average dot diameter characterized by less than 7%variation.
 93. The method of claim 1, wherein the nanomaterial ischaracterized by a bioactivity which is retained upon transport to formthe structure on the surface.
 94. A method comprising simultaneouslypatterning multiple inks from tips to a substrate, wherein the inkscomprise different nanomaterials and polymer, and wherein the differentnanomaterials are transported to the substrate at similar rates becausethe ratios of nanomaterials and polymer in the inks are tuned.
 95. Themethod of claim 94, wherein the nanomaterials are biomolecules.
 96. Themethod of claim 94, wherein the nanomaterials are biomolecules whichretain bioactivity upon patterning.
 97. The method of claim 94, whereinthe nanomaterials are proteins.
 98. The method of claim 94, wherein thepatterning produces dots.
 99. The method of claim 94, wherein thepatterning produces dots with similar diameters.
 100. A nanoarraycomprising: (A) a substrate, and (B) a plurality of patterns on thesubstrate, the patterns being in the form of dots and comprising atleast two different biomolecules in different dots, wherein the dotshave similar sizes.
 101. The nanoarray of claim 100, wherein thenanoarray is produced by simultaneous patterning of the differentbiomolecules.
 102. The nanoarray of claim 100, wherein the biomoleculesretain their bioactivity.